Methods and compositions to treat glycosaminoglycan-associated molecular interactions

ABSTRACT

Therapeutic compounds and methods for inhibiting a glycosaminoglycan (GAG)-associated molecular interaction in a subject, whatever its clinical setting, are described.

RELATED APPLICATIONS

This application is a continuation of U.S. Pat. No. 09/970,148, filedOct. 2, 2001, which is a continuation of U.S. Pat. No. 09/362,505, filedJul. 27, 1999, which claims the benefit of priority under 35 U.S.C.119(e) to copending U.S. Provisional Application 60/094,454, filed onJul. 28, 1998, the entire contents of which is incorporated herein byreference.

BACKGROUND OF THE INVENTION

Glycosaminoglycans (GAGs) have been shown to be involved with the earlysteps of the infectious process associated with several pathogens. Forexample, it is believed that sulfated proteoglycans are used by theinfectious agents as anchors or adsorption moieties for invasion of thehost cells. Several bacterial and viral infectious agents have beenfound to use extracellular membrane components, such as GAGs, to accesshost cells.

Heparan sulfate and/or other sulfated GAGs have been suggested to beinvolved in the infection process by certain bacteria such asStreptococcus pyogenes associated with acute rheumatic fever andpoststreptococcal glomerulonephritis, Chlamydia trachoinatis,Staphylococcus aureus and Pseudomonas aeruginosa (cystic fibrosis),Legionella pneumophila (Legionnaire's disease), Bordetella pertussis(whooping cough), and Mycoplasma pneumoniae. As one example,Streptococcus pyogenes surfaces bind fibronectin, laminin, fibrinogen,nonspecific immunoglobulins A and G, α2-macroglobulin, β2-microglobulinand albumin. Bacterial components do not bind to epithelial orendothelial cells of the kidney but accumulate on the proteoglycan-richregions that connect these cells to the underlying connective tissue.Another example, Chlamydia trachomatis, is one of the most commonsexually transmitted bacterial pathogens in the world. Infection appearsto be facilitated by binding of a heparan sulfate-like GAG present onthe surface of chlamydia, to a heparan sulfate receptor on the targetcell.

Certain types of viri, Herpesviridae, are believed to be associated withHSPG during the infectious process. These viri appear to interact with acells surface through GAGs found on the proteoglycans of the cell plasmamembrane. These GAGs are similar to heparin. Cytomegalovirus (CMV) andHerpes simplex (HSV-1 and HSV-2) are two of the viri which are believedto infect cells via cell surface GAGs.

Although certain agents have been used to suppress infection of hosts bypathogens, there are limitations to their use. For example, thewidespread use of antibiotics has increasingly led to the problem ofresistant pathogens whose growth can no longer be inhibited by knownantibiotics. Thus, the appearance of multi-drug resistant pathogens hasprompted a search for new classes of compounds which are structurallyand/or functionally different from existing drugs. Drugs having newmechanisms of action could be effective against resistant pathogens,where conventional drugs can no longer be used.

SUMMARY OF THE INVENTION

Methods and compositions which are useful in the treatment of conditionsrelated to glycosaminoglycan (GAG)-associated molecular interactions arepresented herein.

In one aspect the invention relates to methods for treating a conditionrelated to a glycosaminoglycan-associated molecular interaction in asubject. The method includes administering to the subject atherapeutically effective amount of a therapeutic compound having theformula:Q

Y⁻X⁺]_(n)   (I)

wherein Y⁻ is an anionic group at physiological pH; Q is a carriermolecule; X⁺ is a cationic group; and n is an integer selected such thatthe biodistribution of the therapeutic compound for an intended targetsite is not prevented while maintaining activity of the therapeuticcompound, or a pharmaceutically acceptable salt or ester thereof, suchthat the glycosaminoglycan-associated molecular interaction is modulatedand the condition is treated. These methods can be used therapeuticallyto treat a subject, e.g., afflicted with a pathogen, or can be usedprophylactically in a subject susceptible to pathogens.

In another embodiment, the therapeutic compound has at least one anionicgroup covalently attached to a carrier molecule. In another embodiment,the anionic group covalently attached to the carrier molecule is asulfonate group. Accordingly, the therapeutic compound can have theformula:Q

SO₃ ⁻X⁺]_(n)   (II)

wherein Q is a carrier molecule; X⁺ is a cationic group; and n is aninteger. In another embodiment, the anionic group is a sulfate group.Accordingly, the therapeutic compound can have the formula:Q

OSO₃ ⁻X⁺]_(n)   (III)wherein Q is a carrier molecule; X⁺ is a cationic group; and n is aninteger. Carrier molecules which can be used include carbohydrates,polymers, peptides, peptide derivatives, aliphatic groups, alicyclicgroups, heterocyclic groups, aromatic groups and combinations thereof.

The invention also provides a method for modulating interactions betweenan infectious agent and a GAG in a subject. The method includesadministering to the subject a therapeutically effective amount of atherapeutic compound having the formula:Q

Y⁻X⁺]_(n)   (I)

wherein Y⁻ is an anionic group at physiological pH; Q is a carriermolecule; X⁺ is a cationic group; and n is an integer selected such thatthe biodistribution of the therapeutic compound for an intended targetsite is not prevented while maintaining activity of the therapeuticcompound, or a pharmaceutically acceptable salt or ester thereof.

In another aspect, methods and therapeutic compositions are providedherein for treating a subject afflicted with a disease, e.g., acuterheumatic fever and poststreptococcal glomerulonephritis, caused byinfection by bacteria such as Streptococcus pyogenes, Chlamydiatrachomatis, Staphylococcus aureus, Pseudomonas aeruginosa, Legionellapneumophila, Bordetella pertussis, and Mycoplasma pneumoniae, such thatthe subject afflicted with the disease is treated. The methods includeadministering to a subject a therapeutically effective amount of atherapeutic compound of formula (I) for treating the infection. Thetherapeutic compound is not carrageenan, pentosan polysulfate, fucoidan,dextran sulfate, heparin, heparan sulfate or dermatan sulfate.

In yet another aspect, the invention provides methods and therapeuticcompositions for treating a subject afflicted with a disease caused byinfection of viri via such as Cytomegalovirus (CMV) and Herpes simplex(HSV-1 and HSV-2), such that the subject afflicted with the disease istreated. The methods include administering to a subject atherapeutically effective amount of a therapeutic compound of formula(I) for treating the disease. The therapeutic compound is not achondroitin sulfate.

In yet a further aspect a packaged pharmaceutical composition fortreating a condition related to a glycosaminoglycan-associated molecularinteraction or for modulating a GAG-associated molecular interaction,e.g., between a GAG and an infectious agent, is described herein. Thepackaged composition includes a container holding a therapeuticallyeffective amount of a pharmaceutical composition for treating thecondition related to a glycosaminoglycan-associated molecularinteraction in a subject. Alternatively, the packaged compositionincludes a container holding a therapeutically effective amount of apharmaceutical composition for modulating a GAG-associated molecularinteraction. The pharmaceutical composition includes at least onetherapeutic compound having the formula:Q

Y⁻X⁺]_(n)   (I)

wherein Y⁻ is an anionic group at physiological pH; Q is a carriermolecule; X⁺ is a cationic group; and n is an integer selected such thatthe biodistribution of the therapeutic compound for an intended targetsite is not prevented while maintaining activity of the therapeuticcompound, or a pharmaceutically acceptable salt or ester thereof.Instructions for using the pharmaceutical composition for treatment ofthe condition related to a glycosaminoglycan-associated molecularinteraction or for modulating the GAG-associated molecular interactionare included in the packaged pharmaceutical composition.

The invention further provides pharmaceutical compositions for treatinga condition related to a glycosaminoglycan-associated molecularinteraction in a subject. Alternatively, the invention providespharmaceutical compositions for modulating a GAG-associated molecularinteraction in a subject. The pharmaceutical compositions include atherapeutically effective amount of a therapeutic compound of theinvention, as described supra, and a pharmaceutically acceptablecarrier.

In further embodiments, the therapeutic compound has at least oneanionic group covalently attached to a carrier molecule. In yet anotherembodiment, the anionic group covalently attached to the carriermolecule is a sulfonate group. Accordingly, the therapeutic compound canhave the formula:Q

SO₃ ⁻X⁺]_(n)   (II)

wherein Q is a carrier molecule; X⁺ is a cationic group; and n is aninteger. In another embodiment, the anionic group is a sulfate group.Accordingly, the therapeutic compound can have the formula:Q

OSO₃ ⁻X⁺]_(n)   (III)

wherein Q is a carrier molecule; X⁺ is a cationic group; and n is aninteger.

BRIEF DESCRIPTION OF THE DRAWING

FIGS. 1-14 depict the chemical structures of compounds described in thespecification.

FIGS. 15-28 illustrate the efficacy of compounds of the invention ininhibiting binding of certain compounds, e.g., Rantes, IL-8, toheparan-coated wells. The compounds referenced in the drawings are: 1)3-amino-1-propanesulfonic acid, sodium salt; 2) trisodiumphosphonoformate; 3) methylene diphosphonic acid; 4) trehaloseoctasulfate, octasodium salt; 5) trans-4-hydroxy-L-proline-4-sulfate,disodium salt; 6) nitrilo(methylene) triphosphonic acid; 7)poly(vinylsulfonate), sodium salt (PVS501, Aldrich); 8)3-[-2-6-methoxy-1,2,3,4-tetrahydroisoquinolinyl)]-1-propanesulfonic add;9) 3-phosphonopropanesulfonic acid, trisodium salt; 10);4,5-dihydroxy-1,3,benzenedisulfonic acid, sodium salt; 11)3-cyclohexylamino-1-propanesulfonic acid; 12) O-phospho-L-serine; and13) 2-thiopheneboronic acid.

DETAILED DESCRIPTION OF THE INVENTION

The features and other details of the invention will now be moreparticularly described with reference to the accompanying drawings andpointed out in the claims. It will be understood that particularembodiments described herein are shown by way of illustration and not aslimitations of the invention. The principal features of this inventioncan be employed in various embodiments without departing from the scopeof the invention. All parts and percentages are by weight unlessotherwise specified.

The invention provides methods and compositions which are useful in thetreatment of conditions related to glycosaminoglycan (GAG)-associatedmolecular interactions. In one embodiment, the invention provides amethod for treating a condition related to aglycosaminoglycan-associated molecular interaction in a subject. Themethod includes administering to the subject, a therapeuticallyeffective amount of a therapeutic compound having the formula:Q

Y⁻X⁺]_(n)   (I)

wherein Y⁻ is an anionic group at physiological pH; Q is a carriermolecule; X⁺ is a cationic group; and n is an integer selected such thatthe biodistribution of the therapeutic compound for an intended targetsite is not prevented while maintaining activity of the therapeuticcompound, or a pharmaceutically acceptable salt of ester thereof. Themethods of the invention can be used therapeutically to treat a subjectafflicted by a pathogen or can be used prophylactically in a subjectsusceptible to pathogens. The methods of the invention are based, atleast in part, on inhibiting, eradicating, or preventing interactionbetween the cell membrane surface and the pathogen.

The language “treating a condition related to a glycosaminoglycan(GAG)-associated molecular interaction” and “treatment of a conditionrelated to a glycosaminoglycan-associated molecular interaction” isintended to include changes in a condition related to aglycosaminoglycan-associated molecular interaction, as described infra,such that physiological symptoms in a subject can be significantlydiminished or minimized. The language also includes control, prevention,relief, or inhibition of physiological symptoms or effects attributed toa disease state associated with glycosaminoglycan-associated molecularinteractions. In one preferred embodiment, the control of theglycosaminoglycan-associated molecular interaction or condition relatedthereto is such that the glycosaminoglycan-associated molecularinteraction or condition related thereto is eradicated. In anotherpreferred embodiment, the control is selective such that a particulartargeted glycosaminoglycan-associated molecular interaction, e.g., witha pathogen, is controlled while other cells and physiological florawhich are not detrimental to the subject are allowed to remainsubstantially uncontrolled or substantially unaffected, e.g.,lymphocytes, red blood cells, white blood cells, platelets, growthfactors, etc.

The term “pathogen” is art recognized and is intended to include diseaseproducing agents, such as organisms, e.g., microorganisms, capable ofcausing disease in a subject, e.g., a mammal, including, for example,bacteria, viruses, prions and fungi.

As used herein, “glycosaminoglycan (GAG)-associated molecularinteraction” is intended to include the binding of a GAG to, forexample, a cell surface, secreted, or extracellular protein. This termalso includes any subsequent results of such protein binding such as,for example, delayed proteolytic degradation or denaturing, changes inprotein conformation (which may, for example, lead to alterations ofbiological activity), or catalysis of a reaction between two differentproteins bound to the same or different GAGs on the same or differentproteoglycans. Also included is the ability of certain GAGs, e.g.,heparin sulfate, to modulate the interaction of a protein to anotherGAG, for example, FGF-2 (basic fibroblast growth factor) to its GAG cellreceptor.

Other GAG-associated molecular interactions include specificinteractions between specific compounds and factors. Non-limitingexamples include polypeptide growth factors (e.g., FGFs1-9, PDGF, HGF,VEGF, TGF-β, IL-3); extracellular matrix components (e.g., laminins,fibronectins; thrombospondins, tenascins, collagens, VonWirebrand'sfactor); proteases and anti-proteases (e.g., thrombin, TPA, UPA, dottingfactors IX and X, PAI-1); cell-adhesion molecules (e.g., N-CAM, LI,myelin-associated glycoprotein); proteins involved in lipoproteinmetabolism (e.g., APO-B, APO-E, lipoprotein lipase); cell-cell adhesionmolecules (e.g., N-CAM, myelin-associated glycoprotein, selectins,pecam); angiogenin; lactoferrin; viral proteins (e.g., proteins fromHIV, herpes complex) and other compounds which bind to GAG. Thedefinition is intended to include the result of the binding of thesefactors to the GAG. For example, the binding of polypeptide growthfactor to a GAG can result in cell proliferation, angiogenesis,inflammation, cancer, and other biologically important responses.

Also, the term “glycosaminoglycan (GAG)-associated molecularinteraction” includes microbial “interactions” with GAGs which may, forexample, lead to invasion of a host cell by a microorganism. It includesthe binding of adhesius or other microbial proteins to GAG and theresults thereafter. Some examples of such adhesius are: the filamentoushemagluggtinin of Bordetella pertussis, gP120 of HIV, gpB and pgC ofHSV, etc. It also includes interactions where the GAG functions as abridge between the microbial organism and the host cell, e.g., inchlamydia trachomatis, heparin sulfate binds to both C. trachomatis tohost cell receptors catalyzing an interaction between the two. Alsoincluded in the definition is any interaction between a microbialorganism and a cell which is mediated through the GAG.

The term “glycosaminoglycan (GAG)-associated molecular interaction” isfurther intended to include disease states or conditions caused by orassociated with one or more pathogens which interact with extracellularmembrane components, e.g., glycosaminoglycans, often found on host cellsurfaces. In one embodiment, the disease state includes, for example,those diseases which afflict a subject by associating with orinterfering with glycosaminoglycans found within the subject. In apreferred embodiment, the term “glycosaminoglycan-associated molecularinteraction” does not include amyloidosis. In another preferredembodiment, the term “glycosaminoglycan-associated molecularinteraction” does not include interactions between an amyloidogenicprotein and a constituent of basement membrane to inhibit amyloiddeposition. In yet another preferred embodiment, the term“glycosaminoglycan” does not include a constituent of basement membrane,e.g., heparan sulfate proteoglycan. In yet another preferred embodiment,the term “glycosaminoglycan” does not include sulfated GAGs, e.g.,heparan sulfate. Presently unknown conditions related toglycosaminoglycan-associated molecular interactions that may bediscovered in the future are encompassed, since their characterizationas conditions related to glycosaminoglycan-associated molecularinteractions will be readily determinable by persons skilled in the art.

Conditions related to glycosaminoglycan-associated molecularinteractions include, for example, certain bacteria such asStreptococcus pyogenes, associated with acute rheumatic fever andpoststreptococcal glomerulonephritis, Chlamydia trachomatis,Staphylococcus aureus (cystic fibrosis), Bordetella pertussis (whoopingcough) and Mycoplasma pneumoniae. For example, Streptococcus pyogenessurfaces bind fibronectin, laminin, fibrinogen, nonspecificimmunoglobulins A and G, α2-macroglobulin, β2-microglobulin and albumin.Infection by Chlamydia trachomatis is facilitated by binding of aheparan sulfate-like GAG present on the surface of chlamydia, to aheparan sulfate receptor on the target cell.

Additionally, conditions related to glycosaminoglycan-associatedmolecular interactions include certain types of viri, such asHerpesviridae, which are believed to be associated with HSPG during theinfectious process. These viri appear to interact with a cell's surfacethrough GAGs found on the proteoglycans of the cell plasma membrane.These GAGs are similar to heparin. Cytomegalovirus (CMV), HIV and Herpessimplex (HSV-1 and HSV-2) are examples of which are believed to infectcells via cell surface GAGs.

In one aspect, the present invention pertains to methods for modulatinga glycosaminoglycan-associated molecular interaction, e.g., between aninfectious agent and a GAG, in a subject. The methods includeadministering to the subject a therapeutically effective amount of atherapeutic compound. The therapeutic compound has the formula:Q

Y⁻X⁺]_(n)   (I)

wherein Y⁻ is an anionic group at physiological pH; Q is a carriermolecule; X⁺ is a cationic group; and n is an integer selected such thatthe biodistribution of the therapeutic compound for an intended targetsite is not prevented while maintaining activity of the therapeuticcompound, or a pharmaceutically acceptable salt or ester thereof.

The term “infectious agent” is intended to include those pathogens whichare associated with disease states caused by bacteria, viri or prions.The term is also intended to include those extracellular components,e.g., proteins, etc., which are secreted, produced, or otherwisedischarged by a pathogen, thereby causing the subject to be afflictedwith a disease state associated with the infectious agent. Those diseasestates associated with infectious agents include Streptococcus pyogenes,Chlamydia trachomatis, Staphylococcus aureus, Bordetella pertussis,Mycoplasma pneumoniae, Herpesvzridae, e.g., herpes simplex. The terminfectious agent is also intended to encompass presently unknowninfectious agents that may be discovered in the future, since theircharacterization as a infectious agents will be readily determinable bypersons skilled in the art.

In an embodiment, therapeutic compounds which comprise at least onesulfate group covalently attached to a carrier molecule, orpharmaceutically acceptable salt thereof are used to treat a conditionrelated to a glycosaminoglycan (GAG)-associated molecular interaction oran infectious agent. In particular, the therapeutic compounds of theinvention comprise at least one sulfate group or a functional equivalentthereof, for example a sulfonic acid group or other functionallyequivalent anionic group, linked to a carrier molecule. In addition tofunctioning as a carrier for the anionic functionality, the carriermolecule can enable the compound to traverse biological membranes and tobe biodistributed without excessive or premature metabolism. Moreover,when multiple anionic functionalities are present on a carrier molecule,the carrier molecule serves to space the anionic groups in a correctgeometric separation.

In one embodiment, when the condition related to aglycosaminoglycan-associated molecular interaction is associated with orcaused by the bacteria Chlamydia trachomatis, the therapeutic compoundis not sulfated polysaccharides kapp and lambda, iota carrageenansC-1263, C-3889 and C-4014, pentosan polysulfate (P-8275), fucodian(F-5631), dextran sulfate (D-6001), heparin (H-3393), heparan sulfate(H-7641) or dermatan sulfate (chondroitin sulfate A and B).

In another embodiment, when the condition related to aglycosaminoglycan-associated molecular interaction is associated with orcaused by cytomegalovirus, the therapeutic compound is not pentosanpolysulfate, dextran sulfate, heparin, copolymers of acrylic acid andvinylalcohol sulfate, α-cyclodextrin hexasulfate and α-cyclodextrindodecasulfate.

In yet another embodiment, when the condition related to aglycosaminoglycan-associated molecular interaction is associated withthe virus Herpesviridae, the therapeutic agent is not chondroitinsulfate A, B or C.

In one embodiment, the method of the invention includes administering tothe subject an effective amount of a therapeutic compound which has atleast one anionic group covalently attached to a carrier molecule. Thetherapeutic compound is capable of treating a condition related to aglycosaminoglycan-associated molecular interaction or an infectiousagent. The therapeutic compound can have the formula:Q

Y⁻X⁺]_(n)   (I)

wherein Y⁻ is an anionic group at physiological pH; Q is a carriermolecule; X⁺ is a cationic group; and n is an integer. The number ofanionic groups (“n”) is selected such that the biodistribution of thecompound for an intended target site is not prevented while maintainingactivity of the compound. For example, the number of anionic groups isnot so great as to inhibit traversal of an anatomical barrier, such as acell membrane, or entry across a physiological barrier, such as theblood-brain barrier, in situations where such properties are desired. Inone embodiment, n is an integer between 1 and 10. In another embodiment,n is an integer between 3 and 8.

An anionic group of a therapeutic compound of the invention is anegatively charged moiety that, when attached to a carrier molecule, caninhibit an interaction between a bacteria, a virus or an infectiousagent and a cell membrane. The anionic group is desirably negativelycharged at physiological pH. Preferably, the anionic therapeuticcompound mimics the structure of a sulfated proteoglycan, i.e., is asulfated compound or a functional equivalent thereof. “Functionalequivalents” of sulfates are intended to include compounds such assulfamates as well as bioisosteres. Bioisosteres encompass bothclassical bioisosteric equivalents and non-classical bioisostericequivalents. Classical and nonclassical bioisosteres of sulfate groupsare known in the art (see, e.g., Silverman, R. B. The Organic Chemistryof Drug Design and Drug Action, Academic Press, Inc. San Diego, Calif.,1992, pp. 19-23). Accordingly, a therapeutic compound of the inventioncan comprise at least one anionic group including sulfonates, sulfates,sulfamates, phosphonates, phosphates, carboxylates, and heterocyclicgroups of the following formulae:

Depending on the carrier molecule, more than one anionic group can beattached thereto. When more than one anionic group is attached to acarrier molecule, the multiple anionic groups can be the same structuralgroup (e.g., all sulfonates) or, alternatively, a combination ofdifferent anionic groups can be used (e.g., sulfonates and sulfates,etc.).

A therapeutic compound of the invention typically further comprises acounter cation (i.e., X⁺ in formula (I): Q

Y⁻X⁺]_(n)). Cationic groups include positively charged atoms andmoieties. If the cationic group is hydrogen, H⁺, then the compound isconsidered an acid, e.g., ethanesulfonic acid. If hydrogen is replacedby a metal or its equivalent, the compound is a salt of the acid.Pharmaceutically acceptable salts of the therapeutic compound are withinthe scope of the invention. For example, X⁺ can be a pharmaceuticallyacceptable alkali metal, alkaline earth, higher valency cation (e.g.,aluminum salt), polycationic counter ion or ammonium. A preferredpharmaceutically acceptable salt is a sodium salt but other salts arealso contemplated within their pharmaceutically acceptable range.

Within the therapeutic compound, the anionic group(s) is covalentlyattached to a carrier molecule. Suitable carrier molecules includecarbohydrates, polymers, peptides, peptide derivatives, aliphaticgroups, alicyclic groups, heterocyclic groups, aromatic groups orcombinations thereof. A carrier molecule can be substituted, e.g. withone or more amino, nitro, halogen, thiol or hydroxy groups.

As used herein, the term “carbohydrate” is intended to includesubstituted and unsubstituted mono-, oligo-, and polysaccharides.Monosaccharides are simple sugars usually of the formula C₆H₁₂O₆ thatcan be combined to form oligosaccharides or polysaccharides.Monosaccharides include enantiomers and both the D and L stereoisomersof monosaccharides. Carbohydrates can have multiple anionic groupsattached to each monosaccharide moiety. For example, in sucroseoctasulfate, four sulfate groups are attached to each of the twomonosaccharide moieties.

As used herein, the term “polymer” is intended to include moleculesformed by the chemical union of two or more combining subunits calledmonomers. Monomers are molecules or compounds which usually containcarbon and are of relatively low molecular weight and simple structure.A monomer can be converted to a polymer by combination with itself orother similar molecules or compounds. A polymer may be composed of asingle identical repeating subunit or multiple different repeatingsubunits (copolymers). Polymers within the scope of this inventioninclude substituted and unsubstituted vinyl, acryl, styrene andcarbohydrate-derived polymers and copolymers and salts thereof. In oneembodiment, the polymer has a molecular weight of approximately 800-1000Daltons. Examples of polymers with suitable covalently attached anionicgroups (e.g., sulfonates or sulfates) includepoly(2-acrylamido-2-methyl-1-propanesulfonic acid);poly(2-acrylamido-2-methyl-1-propanesulfonic acid-co-acrylonitrile);poly(2-acrylamido-2-methyl-1-propanesulfonic acid-co-styrene);poly(vinylsulfonic acid); poly(sodium 4-styrenesulfonic acid); andsulfates and sulfonates derived from: poly(acrylic acid); poly(methylacrylate); poly(methyl methacrylate); and poly(vinyl alcohol); andpharmaceutically acceptable salts thereof. Examples ofcarbohydrate-derived polymers with suitable covalently attached anionicgroups include those of the formula:

wherein R is SO₃ ⁻ or OSO₃ ⁻; and pharmaceutically acceptable saltsthereof.

Peptides and peptide derivatives can also act as carrier molecules. Theterm “peptide” includes two or more amino acids covalently attachedthrough a peptide bond. Amino acids which can be used in peptide carriermolecules include those naturally occurring amino adds found in proteinssuch as glycine, alanine, valine, cysteine, leucine, isoleucine, serine,threonine, methionine, glutamic acid, aspartic acid, glutamine,asparagine, lysine, arginine, proline, histidine, phenylalanine,tyrosine, and tryptophan. The term amino acid further includes analogs,derivatives and congeners of naturally occurring amino acids, one ormore of which can be present in a peptide derivative. For example, aminoadd analogs can have lengthened or shortened side chains or variant sidechains with appropriate functional groups.

Also included are the D and L stereoisomers of an amino acid when thestructure of the amino acid admits of stereoisomeric forms. The term“peptide derivative” further includes compounds which contain moleculeswhich mimic a peptide backbone but are not amino acids (so-calledpeptidomimetics), such as benzodiazepine molecules (see e.g. James, G.L. et al. (1993) Science 260:1937-1942). The anionic groups can beattached to a peptide or peptide derivative through a functional groupon the side chain of certain amino acids or other suitable functionalgroup. For example, a sulfate or sulfonate group can be attached throughthe hydroxy side chain of a serine residue. Accordingly, in oneembodiment, the peptide comprises four amino acids and anionic groups(e.g., sulfonates) are attached to the first, second and fourth aminoacid. For example, the peptide can be Ser-Ser-Y-Ser, wherein an anionicgroup is attached to the side chain of each serine residue and Y is anyamino acid. In addition to peptides and peptide derivatives, singleamino adds can be used as carriers in the therapeutic compounds of theinvention. For example, cysteic acid, the sulfonate derivative ofcysteine, can be used.

The term “aliphatic group” is intended to include organic compoundscharacterized by straight or branched chains, typically having between 1and 22 carbon atoms. Aliphatic groups include alkyl groups, alkenylgroups and alkynyl groups. In complex structures, the chains can bebranched or cross-linked. Alkyl groups include saturated hydrocarbonshaving one or more carbon atoms, including straight-chain alkyl groupsand branched-chain alkyl groups. Such hydrocarbon moieties may besubstituted on one or more carbons with, for example, a halogen, ahydroxyl, a thiol, an amino, an alkoxy, an alkylcarboxy, an alkylthio,or a nitro group. Unless the number of carbons is otherwise specified,“lower aliphatic” as used herein means an aliphatic group, as definedabove (e.g., lower alkyl, lower alkenyl, lower alkynyl), but having fromone to six carbon atoms. Representative of such lower aliphatic groups,e.g., lower alkyl groups, are methyl, ethyl, n-propyl, isopropyl,2-chloropropyl, n-butyl, sec-butyl, 2-aminobutyl, isobutyl, tert-butyl,3-thiopentyl, and the like. As used herein, the term “amino” means —NH₂;the term “nitro” means —NO₂; the term “halogen” designates —F, —Cl, —Bror —I; the term “thiol” means SH; and the term “hydroxyl” means —OH.Thus, the term “alkylamino” as used herein means an alkyl group, asdefined above, having an amino group attached thereto. The term“alkylthio” refers to an alkyl group, as defined above, having asulfhydryl group attached thereto. The term “alkylcarboxyl” as usedherein means an alkyl group, as defined above, having a carboxyl groupattached thereto. The term “alkoxy” as used herein means an alkyl group,as defined above, having an oxygen atom, attached thereto.Representative alkoxy groups include methoxy, ethoxy, propoxy,tert-butoxy and the like. The terms “alkenyl” and “alkynyl” refer tounsaturated aliphatic groups analogous to alkyls, but which contain atleast one double or triple bond respectively.

The term “alicyclic group” is intended to include dosed ring structuresof three or more carbon atoms. Alicyclic groups include cycloparaffinsor naphthenes which are saturated cyclic hydrocarbons, cycloolefinswhich are unsaturated with two or more double bonds, and cycloacetyleneswhich have a triple bond. They do not include aromatic groups. Examplesof cycloparaffins include cyclopropane, cyclohexane, and cyclopentane.Examples of cycloolefins include cyclopentadiene and cyclooctatetraene.Alicyclic groups also include fused ring structures and substitutedalicyclic groups such as alkyl substituted alicyclic groups. In theinstance of the alicyclics such substituents can further comprise alower alkyl, a lower alkenyl, a lower alkoxy, a lower alkylthio, a loweralkylamino, a lower alkylcarboxyl, a nitro, a hydroxyl, —CF₃, —CN, orthe like.

The term “heterocyclic group” is intended to include closed ringstructures in which one or more of the atoms in the ring is an elementother than carbon, for example, nitrogen, or oxygen. Heterocyclic groupscan be saturated or unsaturated and heterocyclic groups such as pyrroleand furan can have aromatic character. They include fused ringstructures such as quinoline and isoquinoline. Other examples ofheterocyclic groups include pyridine and purine. Heterocyclic groups canalso be substituted at one or more constituent atoms with, for example,a halogen, a lower alkyl, a lower alkenyl, a lower alkoxy, a loweralkylthio, a lower alkylamino, a lower alkylcarboxyl, a nitro, ahydroxyl, —CF₃, —CN, or the like.

The term “aromatic group” is intended to include unsaturated cyclichydrocarbons containing one or more rings. Aromatic groups include 5-and 6-membered single-ring groups which may include from zero to fourheteroatoms, for example, benzene, pyrrole, furan, thiophene, imidazole,oxazole, thiazole, triazole, pyrazole, pyridine, pyrazine, pyridazineand pyrimidine, and the like. The aromatic ring may be substituted atone or more ring positions with, for example, a halogen, a lower alkyl,a lower alkenyl, a lower alkoxy, a lower alkylthio, a lower alkylamino,a lower alkylcarboxyl, a nitro, a hydroxyl, —CF₃, —CN, or the like.

The therapeutic compound of the invention can be administered in apharmaceutically acceptable carrier. As used herein “pharmaceuticallyacceptable carrier” includes any and all solvents, dispersion media,coatings, antibacterial and antifungal agents, isotonic and absorptiondelaying agents, and the like which are compatible with the activity ofthe compound and are physiologically acceptable to the subject. Anexample of a pharmaceutically acceptable carrier is buffered normalsaline (0.15 molar NaCl). The use of such media and agents forpharmaceutically active substances is well known in the art. Exceptinsofar as any conventional media or agent is incompatible with thetherapeutic compound, use thereof in the compositions suitable forpharmaceutical administration is contemplated. Supplementary activecompounds can also be incorporated into the compositions.

In an embodiment of the method of the invention, the therapeuticcompound administered to the subject is comprised of at least onesulfonate group covalently attached to a carrier molecule, or apharmaceutically acceptable salt thereof. Accordingly, the therapeuticcompound can have the formula:Q

SO₃ ⁻X⁺]_(n)   (II)

wherein Q is a carrier molecule; X⁺ is a cationic group; and n is aninteger. Suitable carrier molecules and cationic groups are thosedescribed hereinbefore. The number of sulfonate groups (“n”) is selectedsuch that the biodistribution of the compound for an intended targetsite is not prevented while maintaining activity of the compound asdiscussed earlier. In one embodiment, n is an integer between 1 and 10.In another embodiment, n is an integer between 3 and 8. As describedearlier, therapeutic compounds with multiple sulfonate groups can havethe sulfonate groups spaced such that the compound interacts optimallywith a receptor site on the cell membrane.

In certain embodiments, the carrier molecule for a sulfonate(s) is alower aliphatic group (e.g., a lower alkyl, lower alkenyl or loweralkynyl), a heterocyclic group, a disaccharide, a polymer or a peptideor peptide derivative. Furthermore, the carrier can be substituted, e.g.with one or more amino, nitro, halogen, thiol or hydroxy groups. Incertain embodiments, the carrier molecule for a sulfonate(s) is anaromatic group.

Particularly suitable therapeutic compounds include1,3-propanedisulfonic acid, 3-amino-1-propanesulfonic acid,3-dimethylamino-1-propanesulfonic acid sodium salt,2-(3-sulfopropyl)-1,2,3,4-tetrahydro-9H-pyrido[3,4-b]indole, sodiumsalt,3-[2-(6-methoxy-1,2,3,4-tetrahydroisoquinolinyl)]-1-propanesulfonicacid, 3-(2-hydroxyethyl)amino-1-propanesulfonic acid,3-(3-hydroxy-1-propyl)amino-1-propanesulfonic acid,(−)3-[(R)-2-hydroxy-1-propyl]amino-1-propanesulfonic acid,3-(4-hydroxy-1-butyl)amino-1-propanesulfonic acid,3-(5-hydroxy-1-pentyl)amino-1-propanesulfonic acid,3-(6-hydroxy-1-hexyl)amino-1-propanesulfonic acid,3-(2-hydroxyethyl)amino-1-propanesulfonic acid,3-(2-hydroxyethyl)amino-1-propanesulfonic acid,3-(2-hydroxyethyl)amino-1-propanesulfonic acid,3-(2-hydroxyethyl)amino-1-propanesulfonic acid,3-hexylarnino-1-propanesulfonic acid, 3-undecylamino-1-propanesulfonicacid, and 3-octadecylamino-1-propanesulfonic acid, and pharmaceuticallyacceptable salts or esters thereof.

Examples of sulfonated polymeric therapeutic compounds includepoly(2-acrylamido-2-methyl-1-propanesulfonic acid);poly(2-acrylamido-2-methyl-1-propanesulfonic acid-co-acrylonitrile);poly(2-acrylamido-2-methyl-1-propanesulfonic acid-co-styrene);poly(vinylsulfonic acid); poly(sodium 4-styrenesulfonic acid); asulfonic acid derivative of poly(acrylic acid); a sulfonic acidderivative of poly(methyl acrylate); a sulfonic acid derivative ofpoly(methyl methacrylate); and a sulfonate derivative of poly(vinylalcohol); and pharmaceutically acceptable salts thereof.

A suitable sulfonated polymer is poly(vinylsulfonic acid) (PVS) or apharmaceutically acceptable salt thereof, preferably the sodium saltthereof. In one embodiment, PVS having a molecular weight of about800-1000 Daltons is used. PVS may be used as a mixture of stereoisomersor as a single active isomer.

A suitable sulfonated disaccharide is a fully or partially sulfonatedsucrose, or pharmaceutically acceptable salt thereof, such as sucroseoctasulfonate. Other sulfonated saccharides include5-deoxy-1,2-O-isopropylidene-α-D-xylofuranose-5-sulfonic acid (XXIII,shown as the sodium salt).

Suitable lower aliphatic sulfonated compounds for use in the inventioninclude ethanesulfonic acid; 2-aminoethanesulfonic acid (taurine);cysteic acid (3-sulfoalanine or α-amino-β-sulfopropionic acid);1-propanesulfonic acid; 1,2-ethanedisulfonic acid; 1,4-butanedisulfonicacid; 1,5-pentanedisulfonic acid; and 4-hydroxybutane-1-sulfonic acid(VIII, shown as the sodium salt); and pharmaceutically acceptable saltsthereof. Other aliphatic sulfonated compounds contemplated for use inthe invention include 1-butanesulfonic acid (XLVII, shown as the sodiumsalt), 2-propanesulfonic acid (XLIX, shown as the sodium salt),3-pentanesulfonic acid (L, shown as the sodium salt), 4-heptanesulfonicacid (LII, shown as the sodium salt), 1-decanesulfonic acid (XLVIII,shown as the sodium salt); and pharmaceutically acceptable saltsthereof. Sulfonated substituted aliphatic compounds contemplated for usein the invention include 3-amino-1-propanesulfonic acid (XXII, shown asthe sodium salt), 3-hydroxypropanesulfonic acid sulfate (XXXV, shown asthe disodium salt), 1,7-dihydroxy-4-heptanesulfonic acid (LIII, shown asthe sodium salt); and pharmaceutically acceptable salts thereof. Yetother sulfonated compounds contemplated for use in the invention include2-[(4-pyridinyl)amido]ethanesulfonic add (LIV, depicted as the sodiumsalt), and pharmaceutically acceptable salts thereof.

Suitable heterocyclic sulfonated compounds include3-(N-morpholino)propanesulfonic acid; andtetrahydrothiophene-1,1-dioxide-3,4-disulfonic acid; andpharmaceutically acceptable salts thereof.

Aromatic sulfonated compounds include 1,3-benzenedisulfonic acid (XXXVI,shown as the disodium salt), 2,5-dimethoxy-1,4-benzenedisulfonic acid(depicted as the disodium salt, XXXVII, or the dipotassium salt, XXXIX),4-amino-3-hydroxy-1-naphthalenesulfonic add (XLIII),3,4-diamino-1-naphthalenesulfonic acid (XLIV); and pharmaceuticallyacceptable salts thereof.

In another embodiment of the method of the invention, the therapeuticcompound administered to the subject is comprised of at least onesulfate group covalently attached to a carrier molecule, or apharmaceutically acceptable salt thereof. Accordingly, the therapeuticcompound can have the formula:Q

OSO₃ ⁻X⁺]_(n)   (III)

wherein Q is a carrier molecule; X⁺ is a cationic group; and n is aninteger. Suitable carrier molecules and cationic groups are thosedescribed hereinbefore. The number of sulfate groups (“n”) is selectedsuch that the biodistribution of the compound for an intended targetsite is not prevented while maintaining activity of the compound asdiscussed earlier. In one embodiment, n is an integer between 1 and 10.In another embodiment, n is an integer between 3 and 8. As describedearlier, therapeutic compounds with multiple sulfate groups can have thesulfate groups spaced such that the compound interacts optimally with abacteria, virus or an infectious agent or a cell membrane.

In certain embodiments, the carrier molecule for a sulfate(s) is a loweraliphatic group (e.g., a lower alkyl, lower alkenyl or lower alkynyl),an aromatic group, a disaccharide, a polymer or a peptide or peptidederivative. Furthermore, the carrier can be substituted, e.g. with oneor more amino, nitro, halogen, thiol or hydroxy groups.

Examples of sulfated polymeric therapeutic compounds includepoly(2-acrylamido-2-methyl-propyl sulfuric acid);poly(2-acrylamido-2-methyl-propyl sulfuric acid-co-acrylonitrile);poly(2-acrylamido-2-methyl-propyl sulfuric acid-co-styrene);poly(vinylsulfuric acid); poly(sodium 4-styrenesulfate); a sulfatederivative of poly(acrylic acid); a sulfate derivative of poly(methylacrylate); a sulfate derivative of poly(methyl methacrylate); and asulfate derivative of poly(vinyl alcohol); and pharmaceuticallyacceptable salts thereof.

A suitable sulfated polymer is poly(vinylsulfuric acid) orpharmaceutically acceptable salt thereof. A suitable sulfateddisaccharide is sucrose octasulfate or pharmaceutically acceptable saltthereof. Other contemplated sulfated saccharides include the add form ofmethyl-α-D-glucopyranoside 2,3-disulfate (XVI), methyl4,6-O-benzylidene-α-D-glucopyranoside 2,3-disulfate (XVII),2,3,4,3′,4′-sucrose pentasulfate (XXXIII),1,3:4,6-di-O-benzylidene-D-mannitol 2,5-disulfate (XLI), D-mannitol2,5-disulfate (XLII), 2,5-di-O-benzyl-D-mannitol tetrasulfate (XLV); andpharmaceutically acceptable salts thereof.

Suitable lower aliphatic sulfated compounds for use in the inventioninclude ethyl sulfuric acid; 2-aminoethan-1-ol sulfuric acid; 1-propanolsulfuric acid; 1,2-ethanediol disulfuric add; 1,3-propanediol disulfuricacid; 1,4-butanediol disulfuric acid; 1,5-pentanediol disulfuric acid;and 1,4-butanediol monosulfuric acid; and pharmaceutically acceptablesalts thereof. Other sulfated aliphatic compounds contemplated for usein the invention include the acid form of 1,3-cydohexanediol disulfate(XL), 1,3,5-heptanetriol trisulfate (XIX),2-hydroxymethyl-1,3-propanediol trisulfate (XX),2-hydroxymethyl-2-methyl-1,3-propanediol trisulfate (XXI),1,3,5,7-heptanetetraol tetrasulfate (XLVI), 1,3,5,7,9-nonanepentasulfate (LI); and pharmaceutically acceptable salts thereof. Othersulfated compounds contemplated for use in the invention include theacid form of 2-amino-2-hydroxymethyl-1,3-propanediol trisulfate (XXIV),2-benzyloxy-1,3-propanediol disulfate (XXIX), 3-hydroxypropylsulfamicacid sulfate (XXX)2,2′-iminoethanol disulfate (XXXI),N,N-bis(2-hydroxyethyl)sulfamic acid disulfate (XXXII); andpharmaceutically acceptable salts thereof.

Suitable heterocyclic sulfated compounds include 3-(N-morpholino)propanesulfuric acid; and tetrahydrothiophene-1,1-dioxide-3,4-dioldisulfuric acid; and pharmaceutically acceptable salts thereof.

A further aspect of the invention includes pharmaceutical compositionsfor treating conditions related to glycosaminoglycan-associatedmolecular interactions, such as those described supra. The therapeuticcompounds in the methods of the invention, as described hereinbefore,can be incorporated into a pharmaceutical is composition in an amounteffective to treat a condition related to a glycosaminoglycan-associatedmolecular interaction in a pharmaceutically acceptable carrier.

In another embodiment, the pharmaceutical compositions of the inventioninclude a therapeutic compound that has at least one sulfate groupcovalently attached to a carrier molecule, or a pharmaceuticallyacceptable salt thereof, in an amount sufficient to treat a conditionrelated to a glycosaminoglycan-associated molecular interaction, and apharmaceutically acceptable carrier. The therapeutic compound can havethe following formula:Q

OSO₃ ⁻X⁺]_(n)   (III)

wherein Q is a carrier molecule; X⁺ is a cationic group; and n is aninteger selected such that the biodistribution of the compound for anintended target site is not prevented while maintaining activity of thecompound.

The use of prodrugs which are converted in vivo to the therapeuticcompounds of the invention (see, e.g., R. B. Silverman, 1992, “TheOrganic Chemistry of Drug Design and Drug Action”, Academic Press, Chp.8) are also to be considered within the scope of the present invention.Such prodrugs can be used to alter the biodistribution (e.g., to allowcompounds which would not typically cross the blood-brain barrier tocross the blood-brain barrier) or the pharmacokinetics of thetherapeutic compound. For example, an anionic group, e.g., a sulfate orsulfonate, can be esterified, e.g., with a methyl group or a phenylgroup, to yield a sulfate or sulfonate ester. When the sulfate orsulfonate ester is administered to a subject, the ester is cleaved,enzymatically or non-enzymatically, reductively or hydrolytically, toreveal the anionic group. Such an ester can be cyclic, e.g., a cyclicsulfate or sultone, or two or more anionic moieties may be esterifiedthrough a linking group. Exemplary cyclic compounds include, forexample, 2-sulfobenzoic acid (LV), propane sultone (LVI), butane sultone(LVII), 1,3-butanediol cyclic sulfate (LVIII,α-chloro-α-hydroxy-o-toluenesulfonic acid sultone (LIX), and6-nitronaphth-[1,8-cd]-1,2,-oxathiole 2,2-dioxide (LX). In anembodiment, the prodrug is a cyclic sulfate or sultone. An anionic groupcan be esterified with moieties (e.g., acyloxymethyl esters) which arecleaved to reveal an intermediate compound which subsequently decomposesto yield the active compound. In another embodiment, the prodrug is areduced form of a sulfate or sulfonate, e.g., a thiol, which is oxidizedin vivo to the therapeutic compound. Furthermore, an anionic moiety canbe esterified to a group which is actively transported in vivo, or whichis selectively taken up by target organs. The ester can be selected toallow specific targeting of the therapeutic moieties to particularorgans, as described below for carrier moieties.

Carrier molecules useful in the therapeutic compounds include carriermolecules previously described, e.g. carbohydrates, polymers, peptides,peptide derivatives, aliphatic groups, alicyclic groups, heterocyclicgroups, aromatic groups or combinations thereof. Suitable polymersinclude substituted and unsubstituted vinyl, acryl, styrene andcarbohydrate-derived polymers and copolymers and salts thereof. Suitablecarrier molecules include a lower alkyl group, a heterocyclic group, adisaccharide, a polymer or a peptide or peptide derivative.

Carrier molecules useful in the present invention may also includemoieties which allow the therapeutic compound to be selectivelydelivered to a target organ or organs. For example, if delivery of atherapeutic compound to the brain is desired, the carrier molecule mayinclude a moiety capable of targeting the therapeutic compound to thebrain, by either active or passive transport (a “targeting moiety”).Illustratively, the carrier molecule may include a redox moiety, asdescribed in, for example, U.S. Pat. Nos. 4,540,564 and 5,389,623, bothto Bodor. These patents disclose drugs linked to dihydropyridinemoieties which can enter the brain, where they are oxidized to a chargedpyridinium species which is trapped in the brain. Thus, drug accumulatesin the brain. Exemplary pyridine/dihdropyridine compounds of theinvention include sodium 1-(3-sulfopropyl)-1,4-dihydropyridine (LXI),sodium 2-(nicotinylamido)-ethanesulfonate (LXII), and1-(3-sulfopropyl)-pyridinium betaine (LXIII). Other carrier moietiesinclude compounds, such as amino acids or thyroxine, which can bepassively or actively transported in vivo. An illustrative compound isphenylalanyltaurine (LXIX), in which a taurine molecule is conjugated toa phenylalanine (a large neutral amino acid). Such a carrier moiety canbe metabolically removed in vivo, or can remain intact as part of anactive compound. Structural mimics of amino adds (and other activelytransported moieties) are also useful in the invention (e.g.,1-(aminomethyl)-1-(sulfomethyl)-cyclohexane (LXX)). Other exemplaryamino acid mimetics include p(sulfomethyl)phenylalanine (LXXII),p-(1,3-disulfoprop-2-yl)phenylalanine (LXXIII), andO-(1,3-disulfoprop-2-yl)tyrosine (LXXIV). Exemplary thyroxine mimeticsinclude compounds LXXV, LXVI, and LXXVII. Many targeting moieties areknown, and include, for example, asialoglycoproteins (see, e.g. Wu, U.S.Pat. No. 5,166,320) and other ligands which are transported into cellsvia receptor-mediated endocytosis (see below for further examples oftargeting moieties which may be covalently or non-covalently bound to acarrier molecule). Furthermore, the therapeutic compounds of theinvention may bind to bacteria, viri, infectious agents or cellmembranes in the circulation and thus be transported to the site ofaction.

The targeting and prodrug strategies described above can be combined toproduce a compound that can be transported as a prodrug to a desiredsite of action and then unmasked to reveal an active compound. Forexample, the dihydropyrine strategy of Bodor (see supra) can be combinedwith a cyclic prodrug, as for example in the compound2-(1-methyl-1,4-dihydronicotinyl)amidomethyl-propanesultone (LXXI).

In one embodiment, the therapeutic compound in the pharmaceuticalcompositions is a sulfonated polymer, for examplepoly(2-acrylamido-2-methyl-1-propanesulfonic acid);poly(2-acrylamido-2-methyl-1-propanesulfonic acid-co-acrylonitrile);poly(2-acrylamido-2-methyl-1-propanesulfonic acid-co-styrene);poly(vinylsulfonic acid); poly(sodium 4-styrenesulfonic acid); asulfonate derivative of poly(acrylic acid); a sulfonate derivative ofpoly(methyl acrylate); a sulfonate derivative of poly(methylmethacrylate); and a sulfonate derivative of poly(vinyl alcohol); andpharmaceutically acceptable salts thereof.

The therapeutic compound can also have the structure:

in which Z is XR² or R⁴, R¹ and R² are each independently hydrogen, asubstituted or unsubstituted aliphatic group (preferably a branched orstraight-chain aliphatic moiety having from 1 to 24 carbon atoms in thechain; or an unsubstituted or substituted cyclic aliphatic moiety havingfrom 4 to 7 carbon atoms in the aliphatic ring; suitable aliphatic andcyclic aliphatic groups are alkyl groups, more preferably lower alkyl),an aryl group, a heterocyclic group, or a salt-forming cation; R³ ishydrogen, lower alkyl, aryl, or a salt-forming cation; X is,independently for each occurrence, O or S; R⁴ is hydrogen, lower alkyl,aryl or amino; Y¹ and Y² are each independently hydrogen, halogen (e.g.,F, Cl, Br, or I), lower alkyl, amino (including alkylamino,dialkylamino, arylamino, diarylamino, and alkylarylamino), hydroxy,alkoxy, or aryloxy; and n is an integer from 0 to 12 (more preferably 0to 6, more preferably 0 or 1). These compounds are described in U.S.application Ser. No. 08/912,574, the contents of which are incorporatedherein by reference.

Suitable therapeutic compounds for use in the invention includecompounds in which both R¹ and R² are pharmaceutically acceptablesalt-forming cations. It will be appreciated that the stoichiometry ofan anionic compound to a salt-forming counterion (if any) will varydepending on the charge of the anionic portion of the compound (if any)and the charge of the counterion. In a particularly suitable embodiment,R¹, R² and R³ are each independently a sodium, potassium or calciumcation. In certain embodiments in which at least one of R¹ and R² is analiphatic group, the aliphatic group has between 1 and 10 carbons atomsin the straight or branched chain, and is more preferably a lower alkylgroup. In other embodiments in which at least one of R¹ and R² is analiphatic group, the aliphatic group has between 10 and 24 carbons atomsin the straight or branched chain. In certain embodiments, n is 0 or 1;more preferably, n is 0. In certain embodiments of the therapeuticcompounds, Y¹ and Y² are each hydrogen.

In certain embodiments, the therapeutic compound of the invention canhave the structure:

in which R¹, R², R³, Y¹, Y², X and n are as defined above. In otherembodiments, the therapeutic compound of the invention can have thestructure:

in which R¹, R², R³, Y¹, Y², and X are as defined above, R_(a) and R_(b)are each independently hydrogen, alkyl, aryl, or heterocyclyl, or R_(a)and R_(b), taken together with the nitrogen atom to which they areattached, form a cyclic moiety having from 3 to 8 atoms in the ring, andn is an integer from 0 to 6. In certain embodiments, R_(a) and R_(b) areeach hydrogen. In certain embodiments, a compound of the inventioncomprises an α-amino acid (or α-amino acid ester), more preferably aL-α-amino acid or ester.

The Z, Q, R¹, R², R³, Y¹, Y² and X groups are each independentlyselected such that the biodistribution of the therapeutic compound foran intended target site is not prevented while maintaining activity ofthe therapeutic compound. For example, the number of anionic groups (andthe overall charge on the therapeutic compound) should not be so greatas to inhibit traversal of an anatomical barrier, such as a cellmembrane, or entry across a physiological barrier, such as theblood-brain barrier, in situations where such properties are desired.For example, it has been reported that esters of phosphonoformate havebiodistribution properties different from, and in some cases superiorto, the biodistribution properties of phosphonoformate (see, e.g., U.S.Pat. Nos. 4,386,081 and 4,591583 to Helgstrand et al., and U.S. Pat.Nos. 5,194,654 and 5,463,092 to Hostetler et al.). Thus, in certainembodiments, at least one of R¹ and R² is an aliphatic group (morepreferably an alkyl group), in which the aliphatic group has between 10and 24 carbons atoms in the straight or branched chain. The number,length, and degree of branching of the aliphatic chains can be selectedto provide a desired characteristic, e.g., lipophilicity. In otherembodiments, at least one of R¹ and R² is an aliphatic group (morepreferably an alkyl group), in which the aliphatic group has between 1and 10 carbons atoms in the straight or branched chain. Again, thenumber, length, and degree of branching of the aliphatic chains can beselected to provide a desired characteristic, e.g., lipophilicity orease of ester cleavage by enzymes. In certain embodiments, a suitablealiphatic group is an ethyl group.

In another embodiment, the therapeutic compound of the invention canhave the structure:

in which G represents hydrogen or one or more substituents on the arylring (e.g., alkyl, aryl, halogen, amino, and the like) and L is asubstituted alkyl group (in certain embodiments, preferably a loweralkyl), more preferably a hydroxy-substituted alkyl or an alkylsubstituted with a nucleoside base. In certain embodiments, G ishydrogen or an electron-donating group. In embodiments in which G is anelectron-withdrawing group, G is preferably an electron withdrawinggroup at the meta position. The term “electron-withdrawing group” isknown in the art, and, as used herein, refers to a group which has agreater electron-withdrawing than hydrogen. A variety ofelectron-withdrawing groups are known, and include halogens (e.g.,fluoro, chloro, bromo, and iodo groups), nitro, cyano, and the like.Similarly, the term “electron-donating group”, as used herein, refers toa group which is less electron-withdrawing than hydrogen. In embodimentsin which G is an electron donating group, G can be in the ortho, meta orpara position.

In certain embodiments, L is a moiety selected from the group consistingof:

Table 1 lists data pertinent to the characterization of these compoundsusing art-recognized techniques. TABLE 1 COMPOUND ³¹P NMR ¹³C NMRFAB-MS(−) IVa −6.33(DMSO-d₆) 60.97 CH₂OH(d, J=6Hz) 245.2 66.76 CHOH(d,J=7.8Hz) 121.65, 121.78, 121.99, 125.71, 129.48, 129.57, 126.43 AromaticCH 134.38 Aniline C—N 150.39 Phenyl C—O(d, J=7Hz) 171.57 P—C═O(d,J=234Hz) IVb −6.41(DMSO-d₆) 13.94 CH₃ 456 22.11, 24.40, 28.56, 28.72,28.99, 29.00, 31.30, 33.43, —(CH₂)₁₀— 65.03 CH₂—OC(O) 66.60 CH₂—OP(d,J=5.6Hz) 67.71 CH2-OH(d, J=6Hz) 121.73, 121.10, 125.64, 126.57, 129.40,129.95, Aromatic CH 134.04 Aniline C—N 150.31 Phenyl C—O 171.44 P—C═O(d,J=6.7Hz) 172.83 O—C═O IVc −6.46(DMSO-d₆) 13.94 CH₃ 471 22.11, 25.10,28.68, 28.72, 28.85, 29.00, 30.76, 31.31, 32.10, —(CH₂)₁₀— 43.36 CH₂—S68.43 CH₂—OH 68.43 CH—OH(d, J=6.3Hz) 68.76 P—O—CH₂-9d, J=5.8Hz) 121.75,122.03, 125.62, 126.37, 129.30, 129.53, Aromatic CH 134.23 Aniline C—N150.37 Phenyl C—O(d, J=6.7Hz) 171.47 P—C═O(d, J=234.0Hz) 198.47 S—C═OIVd −6.61(DMSO-d₆₎ _(13.94) CH₃ 416 22.06, 25.14, 28.24, 28.35, 31.09,32.14 —CH₂)₆₋ 43.40 CH₂—S 68.50 P—O—CH₂—(d, J=5.8Hz) 68.77 CH—OH(d,6.4Hz) 121.78, 122.59, 125.69, 127.06, 129.43, 129.59 Aromatic CH 133.39Aniline C—N 150.38 Phenyl C—O(d, J=6.7Hz) 171.47 P—C═O(d, J=234.4Hz)198.54 S—C═O IVe −5.76(D₂O) N/A N/A IVf −7.00(DMSO-d₆) N/A N/A IVg−6.60(DMSO-D6) 70.84 CH2-OH 321 72.17 CH—OH 121.68, 121.79, 121.85,125.71 127.10, 127.92, 129.36, 129.50, 129.59 Aromatic CH 134.51 AnilineC—N 142.34 Aromatic C—CH 150.37 Phenyl C—O(d, J=6.2Hz) 171.59 P—C═O(d,J=232.6Hz)

It will be noted that the structure of some of the therapeutic compoundsof this invention includes asymmetric carbon atoms. It is to beunderstood accordingly that the isomers (e.g., enantiomers anddiastereomers) arising from such asymmetry are included within the scopeof this invention. Such isomers can be obtained in substantially pureform by classical separation techniques and by sterically controlledsynthesis. For the purposes of this application, unless expressly notedto the contrary, a therapeutic compound shall be construed to includeboth the R or S stereoisomers at each chiral center.

In certain embodiments, an therapeutic compound of the inventioncomprises a cation (i.e., in certain embodiments, at least one of R¹, R²or R³ is a cation). If the cationic group is hydrogen, H⁺, then thetherapeutic compound is considered an acid, e.g., phosphonoformic acid.If hydrogen is replaced by a metal ion or its equivalent, thetherapeutic compound is a salt of the acid. Pharmaceutically acceptablesalts of the therapeutic compound are within the scope of the invention.For example, at least one of R¹, R² or R³ can be a pharmaceuticallyacceptable alkali metal (e.g., Li, Na, or K), ammonium cation, alkalineearth cation (e.g., Ca²⁺, Ba²⁺, Mg²⁺), higher valency cation, orpolycationic counter ion (e.g., a polyammonium cation). (See, e.g.,Berge et al. (1977) “Pharmaceutical Salts”, J. Pharm. Sci. 66:1-19). Itwill be appreciated that the stoichiometry of an anionic compound to asalt-forming counterion (if any) will vary depending on the charge ofthe anionic portion of the compound (if any) and the charge of thecounterion. Preferred pharmaceutically acceptable salts include asodium, potassium or calcium salt, but other salts are also contemplatedwithin their is pharmaceutically acceptable range.

The term “pharmaceutically acceptable esters” refers to the relativelynon-toxic, esterified products of the therapeutic compounds of thepresent invention. These esters can be prepared in situ during the finalisolation and purification of the therapeutic compounds or by separatelyreacting the purified therapeutic compound in its free acid form orhydroxyl with a suitable esterifying agent; either of which are methodsknown to those skilled in the art Carboxylic acids and phosphonic acidscan be converted into esters according to methods well known to one ofordinary skill in the art, e.g., via treatment with an alcohol in thepresence of a catalyst. A preferred ester group (e.g., when R³ is loweralkyl) is an ethyl ester group.

The term “alkyl” refers to the saturated aliphatic groups, indudingstraight-chain alkyl groups, branched-chain alkyl groups, cycloalkyl(alicyclic) groups, alkyl substituted cydoalkyl groups, and cycloalkylsubstituted alkyl groups. In preferred embodiments, a straight chain orbranched chain alkyl has 30 or fewer carbon atoms in its backbone (e.g.,C₁-C₃₀ for straight chain, C₃-C₃₀ for branched chain), and morepreferably 20 or fewer. Likewise, preferred cycloalkls have from 4-10carbon atoms in their ring structure, and more preferably have 4-7carbon atoms in the ring structure. The term “lower alkyl” refers toalkyl groups having from 1 to 6 carbons in the chain, and to cycloalklshaving from 3 to 6 carbons in the ring structure.

Moreover, the term “alkyl” (including “lower alkyl”) as used throughoutthe specification and claims is intended to include both “unsubstitutedalkyls” and “substituted alkyls”, the latter of which refers to alkylmoieties having substituents replacing a hydrogen on one or more carbonsof the hydrocarbon backbone. Such substituents can include, for example,halogen, hydroxyl, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy,aryloxycarbonyloxy, carboxylate, alkylcarbonyl, alkoxycarbonyl,aminocarbonyl, alkylthiocarbonyl, alkoxyl, phosphate, phosphonato,phosphinato, cyano, amino (including alkyl amino, dialkylamino,arylamino, diarylamino, and alkylarylamino), acylamino (includingalkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido), amidino,imino, sulfhydryl, alkylthio, arylthio, thiocarboxylate, sulfate,sulfonato, sulfamoyl, sulfonamido, nitro, trifluoromethyl, cyano, azido,heterocyclyl, aralkyl, or an aromatic or heteroaromatic moiety. It willbe understood by those skilled in the art that the moieties substitutedon the hydrocarbon chain can themselves be substituted, if appropriate.Cycloalkyls can be further substituted, e.g., with the substituentsdescribed above. An “aralkyl” moiety is an alkyl substituted with anaryl (e.g., phenylmethyl(benzyl)).

The term “alkoxy”, as used herein, refers to a moiety having thestructure —O-alkyl, in which the alkyl moiety is described above.

The term “aryl” as used herein includes 5- and 6-membered single-ringaromatic groups that may include from zero to four heteroatoms, forexample, unsubstituted or substituted benzene, pyrrole, furan,thiophene, imidazole, oxazole, thiazole, triazole, pyrazole, pyridine,pyrazine, pyridazine and pyrimidine, and the like. Aryl groups alsoinclude polycyclic fused aromatic groups such as naphthyl, quinolyl,indolyl, and the like. The aromatic ring can be substituted at one ormore ring positions with such substituents, e.g., as described above foralkyl groups. Preferred aryl groups include unsubstituted andsubstituted phenyl groups.

The term “aryloxy”, as used herein, refers to a group having thestructure —O-aryl, in which the aryl moiety is as defined above.

The term “amino,” as used herein, refers to an unsubstituted orsubstituted moiety of the formula —NR_(a)R_(b), in which R_(a) and R_(b)are each independently hydrogen, alkyl, aryl, or heterocyclyl, or R_(a)and R_(b), taken together with the nitrogen atom to which they areattached, form a cyclic moiety having from 3 to 8 atoms in the ring.Thus, the term “amino” is intended to include cyclic amino moieties suchas piperidinyl or pyrrolidinyl groups, unless otherwise stated. An“amino-substituted amino group” refers to an amino group in which atleast one of R_(a) and R_(b), is further substituted with an aminogroup.

In another embodiment, R¹ or R² can be (for at least one occurrence) along-chain aliphatic moiety. The term “long-chain aliphatic moiety” asused herein, refers to a moiety having a straight or branched chainaliphatic moiety (e.g., an alkyl or alkenyl moiety) having from 10 to 24carbons in the aliphatic chain, e.g., the long-chain aliphatic moiety isan aliphatic chain of a fatty acid (preferably a naturally-occurringfatty acid). Representative long-chain aliphatic moieties include thealiphatic chains of stearic acid, oleic acid, linolenic add, and thelike.

In certain embodiments, the therapeutic compound of the invention canhave the structure:

in which R¹ and R² are each independently hydrogen, an aliphatic group(preferably a branched or straight-chain aliphatic moiety having from 1to 24 carbon atoms, more preferably 10-24 carbon atoms, in the chain; oran unsubstituted or substituted cyciic aliphatic moiety having from 4 to7 carbon atoms in the aliphatic ring), an aryl group, a heterocyclicgroup, or a salt-forming cation; R³ is hydrogen, lower alkyl, aryl, or asalt-forming cation; Y¹ and Y² are each independently hydrogen, halogen(e.g., F, Cl, Br, or I), lower alkyl, hydroxy, alkoxy, or aryloxy; and nis an integer from 0 to 12. Suitable therapeutic compounds for use inthe invention include compounds in which both R¹ and R² arepharmaceutically acceptable salt-forming cations. In a particularlysuitable embodiment, R¹, R² and R³ are each independently a sodium,potassium or calcium cation, and n is 0. In certain embodiments of thetherapeutic compounds, Y¹ and Y² are each hydrogen. Suitable therapeuticcompounds include salts of phosphonoformate. Trisodium phosphonoformate(foscarnet sodium or Foscavir®) is commercially available (e.g., fromAstra), and its clinical pharmacology has been investigated (see, e.g.,“Physician's Desk Reference”, 51st Ed., pp. 541-545 (1997)).

In another embodiment, the therapeutic compound used in the inventioncan be an aminophosphonate, a biphosphonate, a phosphonocarboxylatederivative, a phosphonate derivative, or a phosphono carbohydrate. Forexample, the therapeutic compound can be one of the compounds describedin Appendix A submitted herewith.

Suitable therapeutic compounds for inclusion in a pharmaceuticalcomposition for treating glycosaminoglycan-associated molecularinteractions include 1,3-propanedisulfonic acid,3-amino-1-propanesulfonic acid, 3-dimethylamino-1-propanesulfonic acidsodium salt,2-(3-sulfopropyl)-1,2,3,4-tetrahydro-9H-pyrido[3,4-b]indole, sodiumsalt,3-[2-(6-methoxy-1,2,3,4-tetrahydroisoquinolinyl)]-1-propanesulfonicacid, 3-(2-hydroxyethyl)amino-1-propanesulfonic add,3-(3-hydroxy-1-propyl)amino-1-propanesulfonic acid,(−)3-[(R)-2-hydroxy-1-propyl]amino-1-propanesulfonic acid,3-(4-hydroxy-1-butyl)amino-1-propanesulfonic acid,3-(5-hydroxy-1-pentyl)amino-1-propanesulfonic acid,3-(6-hydroxy-1-hexyl)amino-1-propanesulfonic acid,3-(2-hydroxyethyl)amino-1-propanesulfonic acid,3-(2-hydroxyethyl)amino-1-propanesulfonic acid,3-(2-hydroxyethyl)amino-1-propanesulfonic acid,3-(2-hydroxyethyl)amino-1-propanesulfonic acid,3-hexylamino-1-propanesulfonic acid, 3-undecylamino-1-propanesulfonicacid, and 3-octadecylamino-1-propanesulfonic acid, and pharmaceuticallyacceptable salts or esters thereof.

In the methods of the invention, a condition related to aglycosaminoglycan-associated molecular interaction in a subject istreated by administering a therapeutic compound of the invention to thesubject. The term “subject” is intended to include living organisms inwhich conditions related to glycosaminoglycan-associated molecularinteractions can occur. The term subject is also intended to includethose living organisms which are afflicted by infectious agents whichsecrete components which interfere with a host cells viaglycosaminoglycans. Examples of subjects include humans, monkeys, cows,sheep, goats, dogs, cats, mice, rats, and transgenic species thereof.Administration of the compositions of the present invention to a subjectto be treated can be carried out using known procedures, at dosages andfor periods of time effective to treat a condition related to aglycosaminoglycan-associated molecular interaction in the subject. Aneffective amount of the therapeutic compound necessary to achieve atherapeutic effect may vary according to factors such as the age, sex,and weight of the subject, and the ability of the therapeutic compoundto treat the foreign agents in the subject. Dosage regimens can beadjusted to provide the optimum therapeutic response. For example,several divided doses may be administered daily or the dose may beproportionally reduced as indicated by the exigencies of the therapeuticsituation. A non-limiting example of an effective dose range for atherapeutic compound of the invention (e.g., poly(vinylsulfonate sodiumsalt)) is between 5 and 500 mg/kg of body weight/per day. In an aqueouscomposition, suitable concentrations for the active compound (i.e., thetherapeutic compound that can treat the disease) are between 5 and 500mM, between 10 and 100 mM, and between 20 and 50 mM.

The therapeutic compounds of the invention may be administered orally.Alternatively, the active compound may be administered by other suitableroutes such subcutaneous, intravenous, intraperitoneal, etc.administration (e.g. by injection). Depending on the route ofadministration, the active compound may be coated in a material toprotect the compound from the action of acids and other naturalconditions which may inactivate the compound.

The compounds of the invention can be formulated to ensure properdistribution in vivo. For example, the blood-brain barrier (BBB) exdudesmany highly hydrophilic compounds. To ensure that the therapeuticcompounds of the invention cross the BBB, they can be formulated, forexample, in liposomes. For methods of manufacturing liposomes, see,e.g., U.S. Pat. Nos. 4,522,811; 5,374,548; and 5,399,331. The liposomesmay comprise one or more moieties which are selectively transported intospecific cells or organs (“targeting moieties”), thus providing targeteddrug delivery (see, e.g., V. V. Ranade (1989) J. Clin. Pharmacol.29:685). Exemplary targeting moieties include folate or biotin (see,e.g., U.S. Pat. No. 5,416,016 to Low et al.); mannosides (Umezawa etal., (1988) Biochem. Biophys. Res. Commun. 153:1038); antibodies (P. G.Bloeman et al. (1995) FEBS Lett. 357:140; M. Owais et al. (1995)Antimicrob. Agents Chemother. 39:180); surfactant protein A receptor(Briscoe et al. (1995) Am. J. Physiol. 1233:134); gp120 (Schreier et al.(1994) J. Biol. Chem. 269:9090); see also K. Keinanen; M. L. Laukkanen(1994) FEBS Lett. 346:123; J. J. Killion; I. J. Fidler (1994)Immunomethods 4:273. In an embodiment, the therapeutic compounds of theinvention are formulated in liposomes; in a more preferred embodiment,the liposomes include a targeting moiety.

Delivery and in vivo distribution can also be affected by alteration ofan anionic group of compounds of the invention. For example, anionicgroups such as carboxylate or tetrazole can be employed instead of, orin addition to, sulfate or sulfonate moieties, to provide compounds withdesirable pharmocokinetic, pharmacodynamic, biodistributive, or otherproperties. Exemplary tetrazole-substituted compounds include3-(1H-tetrazol-5-yl)-9H-thioxanthen-9-one 10,10-dioxide (LXIV),5,5-dithiobis(1-phenyltetrazole) (LXV), 1H-tetrazole (LXVI),5-phenyl-1H-tetrazole (LXVII), and 5-(2-aminoethanoic acid)-1H-tetrazole(LXVIII), and the like; and their pharmaceutically acceptable salts.Exemplary carboxylate-substituted compounds include dicarboxylic acidssuch as adipic add, azelaic acid, 3,3-dimethylglutaric acid, subericacid, succinic acid, and the like, and their pharmaceutically acceptablesalts.

To administer the therapeutic compound by other than parenteraladministration, it may be necessary to coat the compound with, orco-administer the compound with, a material to prevent its inactivation.For example, the therapeutic compound may be administered to a subjectin an appropriate carrier, for example, liposomes, or a diluent.Pharmaceutically acceptable diluents include saline and aqueous buffersolutions. Liposomes include water-in-oil-in-water CGF emulsions as wellas conventional liposomes (Strejan et al., (1984) J. Neuroimmunol.7:27).

The therapeutic compound may also be administered parenterally,intraperitoneally, intraspinally, or intracerebrally. Dispersions can beprepared in glycerol, liquid polyethylene glycols, and mixtures thereofand in oils. Under ordinary conditions of storage and use, thesepreparations may contain a preservative to prevent the growth ofmicroorganisms.

Pharmaceutical compositions suitable for injectable use include sterileaqueous solutions (where water soluble) or dispersions and sterilepowders for the extemporaneous preparation of sterile injectablesolutions or dispersion. In all cases, the composition must be sterileand must be fluid to the extent that easy syringability exists. It mustbe stable under the conditions of manufacture and storage and must bepreserved against the contaminating action of microorganisms such asbacteria and fungi. The carrier can be a solvent or dispersion mediumcontaining, for example, water, ethanol, polyol (for example, glycerol,propylene glycol, and liquid polyethylene glycol, and the like),suitable mixtures thereof, and vegetable oils. The proper fluidity canbe maintained, for example, by the use of a coating such as lecithin, bythe maintenance of the required particle size in the case of dispersionand by the use of surfactants. Prevention of the action ofmicroorganisms can be achieved by various antibacterial and antifungalagents, for example, parabens, chlorobutanol, phenol, ascorbic acid,thimerosal, and the like. In many cases, it will be preferable toinclude isotonic agents, for example, sugars, sodium chloride, orpolyalcohols such as mannitol and sorbitol, in the composition.Prolonged absorption of the injectable compositions can be brought aboutby including in the composition an agent which delays absorption, forexample, aluminum monostearate or gelatin.

Sterile injectable solutions can be prepared by incorporating thetherapeutic compound in the required amount in an appropriate solventwith one or a combination of ingredients enumerated above, as required,followed by filtered sterilization. Generally, dispersions are preparedby incorporating the therapeutic compound into a sterile carrier whichcontains a basic dispersion medium and the required other ingredientsfrom those enumerated above. In the case of sterile powders for thepreparation of sterile injectable solutions, the preferred methods ofpreparation are vacuum drying and freeze-drying which yields a powder ofthe active ingredient (i.e., the therapeutic compound) plus anyadditional desired ingredient from a previously sterile-filteredsolution thereof.

The therapeutic compound can be orally administered, for example, withan inert diluent or an assimilable edible carrier. The therapeuticcompound and other ingredients may also be enclosed in a hard or softshell gelatin capsule, compressed into tablets, or incorporated directlyinto the subject's diet. For oral therapeutic administration, thetherapeutic compound may be incorporated with excipients and used in theform of ingestible tablets, buccal tablets, troches, capsules, elixirs,suspensions, syrups, wafers, and the like. The percentage of thetherapeutic compound in the compositions and preparations may, ofcourse, be varied. The amount of the therapeutic compound in suchtherapeutically useful compositions is such that a suitable dosage willbe obtained.

It is especially advantageous to formulate parenteral compositions indosage unit form for ease of administration and uniformity of dosage.Dosage unit form as used herein refers to physically discrete unitssuited as unitary dosages for the subjects to be treated; each unitcontaining a predetermined quantity of therapeutic compound calculatedto produce the desired therapeutic effect in association with therequired pharmaceutical carrier. The specification for the dosage unitforms of the invention are dictated by and directly dependent on (a) theunique characteristics of the therapeutic compound and the particulartherapeutic effect to be achieved, and (b) the limitations inherent inthe art of compounding such a therapeutic compound for the treatment ofa condition related to a glycosaminoglycan-associated molecularinteraction in a subject.

Active compounds are administered at a therapeutically effective dosagesufficient to treat a condition related to aglycosaminoglycan-associated molecular interaction in a subject. A“therapeutically effective dosage” preferably reduces the amount ofsymptoms of the condition in the infected subject by at least about 20%,more preferably by at least about 40%, even more preferably by at leastabout 60%, and still more preferably by at least about 80% relative tountreated subjects. For example, the ability of a compound to reduce aninfectious agent can be evaluated in an animal model system that may bepredictive of efficacy in treating diseases associated with theinfectious agent in humans.

The invention is further illustrated by the following Exemplification,which should not be construed as further limiting the subject invention.The contents of all references, issued patents, and published patentapplications cited throughout this application including the backgroundare hereby incorporated by reference.

EXEMPLIFICATION

Bacteria:

Streptococcus pyogens:

S. pyogenes can cause acute rheumatic fever streptococcus and acutepost-streptococcal glomerulonephritis. Studies have identified theprotein responsible for stabilizing the bacteria on the basal laminae ofcardiac muscle as well as on kidney tissues.

The mechanism for such a bacterial virulence is unknown. One hypothesisis the direct binding of bacterial adhesions and exotoxins to thecardiac muscle and kidney_Bergey & Stenson identified two streptococcalproteins (9.15 Kda) that are capable of binding to basement membranes ofcardiac muscle and renal tissues; the binding was completely inhibitedby heparin and other GAGs. Binding of specific S. pyogenes protein canbe measured on cardiac muscle section and kidney section. Briefly, S.pyogenes protein preparation are incubated on cardiac muscle and kidneypreparation. Binding of protein is visualized by indirectimmunofluorescence using an antibody against the bacteria protein.Ability of a compound to interfere in such a binding can be determinedin ligand inhibition studies. Binding is measured by comparing theamount of protein binding (as determined by Image Analysis) in presenceor absence of the compound. Measurement can also be determined using3H-labeled streptococcal protein.

Fluorescence: Cryostat-cut section of heart tissue are pre-incubatedwith bacterial antigen preparation. Amount of peptide binding to tissueis determined by indirect immunofluorescence using a Rabbit anti-S.pyogenes serum. Ability of a compound to inhibit the binding of thestreptococcal protein to the tissue is evaluated by comparing the amountof bacterial antigen present of tissue section in presence or absence ofthe inhibitor.

Direct Binding Assay: Radiolabefled streptococcal components are testedfor direct binding activity to mammalian tissue component as previouslydescribed (M W Stinson & E J Bergey, 1982). Dried cardiac musclefragments are rehydrated with PBS and 1% bovine serum albumin. Moistheart material is incubated with radiolabelled bacterial components.Bound radioactivity is determined by liquid scintillation spectrometry.

Determination of a compound's ability to inhibit this binding is donewith various concentrations of the compound added to the streptococcalpreparation prior its incubation with the cardiac muscle preparation.

S. aureus, Pseudomonas aeruglnosa, Legionella pneumophila

S. aureus and P. aeruginosa are well-known to cause major pulmonaryinfection in patients with cystic fibrosis. Legionella pneumophila isknown to cause Legionnaire's disease in susceptible individuals. Thesebacteria need to adhere to mucus membrane to in order to multiply andcause infection.

The ability of specific compounds to inhibit S. Aureus, P. Aeruginosaand L. pneumophila adherence to mucosal membrane can be determined invitro using murine trachea culture. The number of bacteria adhering tothe preparation can be determined by comparing the number of bacteriaremaining in supernatant after incubation with trachea preparation.Briefly, trachea preparation are incubated with a bacterial suspensionin presence or absence of a compound. 30 minutes later the amount ofbacteria remaining the supernatant (i.e., non-adhering) determined byserial dilution.

The ability of bacteria to infect cells an also be determined in vitro.Macrophages are incubated with bacteria and the phagocytic rate isdetermined 30 minutes later.

In vivo: Intratracheal infection with P. aeruginosa or S. aureus with orwithout treatment with compound. Intratracheal infection with P.aeruginosa, S. aureus and Legionella have been shown to cause acutepulmonary infection in mice. The ability of a compound to inhibit suchan infection can be determined by evaluating the bacterial load presentin the lung of infected mice undergoing a treatment with specificcompounds. These compounds can be administered IV, PO, or under aerosol.

Viral Infections:

The infectious process of viruses of the herpesviridae family have beenextensively studied. It has been established that the initialinteraction of several herpes viruses with the cell surface is mediatedby glycosaminoglycans found on the proteoglycans in the cell plasmamembrane. These GAGs are similar to heparin. Amongst the differentherpes viruses found to interact with cell surface GAGs, interestingones are Cytomegalovirus (CMV) and Herpes simplex (HSV-1 and HSV-2). Theability of compounds to interfere in the infectious process of theseviruses is determined as follows:

In vitro: Hela cells are infected with CMV or HSV-1 in presence orabsence of compounds. Ability of CMV to infect cells is determined byevaluating virus load 24-72 hours later by:

-   -   % viral antigen expression (IF)    -   specific viral antigen (mRNA level)    -   Virus particle titration    -   cytopathic effect

The ability of a compound to interfere in the infectious process can bedetermined by evaluating (by the different techniques mentioned above)the amount of virus found in the culture in presence or absence of aninhibitor.

EQUIVALENTS

Those skilled in the art will recognize, or be able to ascertain usingno more than routine experimentation, numerous equivalents to thespecific procedures described herein. Such equivalents are considered tobe within the scope of this invention and are covered by the followingclaims.

APPENDIX A

AMINOPHOSPHONATES Name Structure3-[2-(1,2,3,4-Tetrahydroisoquinolinyl)]-1-pro- panephosphonic acid,disodium salt

3-Aminopropylphosphonic acid NH₂CH₂CH₂CH₂PO₃H₂(S)-2-Amino-2-methyl-4-phos- phonobutanoic acid

D-(−)-2-Amino-4-phosphonobutanoic acid

L-(+)-2-Amino-4-phosphonobutanoic acid

3-Aminopropyl(methyl)phosphinic acid, hydrochloride

(R)-(−)-3-(2-Carboxypiperazin-4-yl)-pro- pyl-1-phosphonic acid(D-CPP)

(R,E)-4-(3-Phosphonoprop-2-en- yl)piperazine-2-carboxylic acid

trans-L-4-Phosphonomethylproline, trisodium salt

cis-L-4-Phosphonomethylproline, trisodium salt

4-Amino-1-butylphosphonic acid, disodium salt

1-(3-Phosphonopropyl)-benzimidazole, disodium salt

3-Dimethylamino-1-propylphosphonic acid, disodium salt

3-Amino-butylphosphonic acid, disodium salt

3-Amino-pentylphosphonic acid, disodium salt

3-Amino-hexylphosphonic acid, disodium salt

3-Amino-heptylphosphonic acid, disodium salt

3-Amino-octylphophonic acid, disodium salt

3-Amino-4-methyl-pentylphosphonic acid, disodium salt

3-Amino-3-methyl-butylphosphonic acid, disodium salt

3-Amino-3-phenyl-propylphosphonic acid, disodium salt

3-Amino-4-phenyl-butylphosphonic acid, disodium salt

3-Amino-4-phenyl-pentylphosphonic acid, disodium salt

3-Amino-3-phenyl-butylphosphonic acid, disodium salt

2-Amino-2-(2-phosphonoethyl)-1,3,4-tri- hydronaphthalene, disodium salt

1-Amino-1-(2-phosphonoethyl)-cyclo- hexane, disodium salt

2-(2-Amino-4-phos- phonobutoxy)tetrahydropyran

3-Amino-4-hydroxy-butylphosphonic acid, disodium salt

Diethyl 2-pyrrolidinylphosphonate

2-Pyrrolidinylphosphonic acid, disodium salt

1,1-Dioxo-2-(3-phosphonopropyl)-iso- thiazoline, disodium salt

2-Deoxy-2-phosphonoacetylamino-D-glucose

3-Hydroxy-3-(2-pyridyl)propenyl-2-phos- phonic acid, disodium salt

3-Hydroxy-3-(3-pyridyl)propenyl-2-phos- phonic acid, disodium salt

3-Hydroxy-3-(4-pyridyl)propenyl-2-phos- phonic acid, disodium salt

3-Amino-3-(2-pyridyl)propenyl-2-phos- phonic acid, disodium salt

3-Amino-3-(3-pyridyl)propenyl-2-phos- phonic acid, disodium salt

3-Amino-3-(4-pyridyl)propenyl-2-phos- phonic acid, disodium salt

1,4-Diamino-1-(3-pyridyl)butyl-2-phos- phonic acid, disodium salt

1,4-Diamino-4-methyl-1-(3-py- ridyl)pentyl-2-phosphonic acid, disodiumsalt

1,4-Diamino-4-methyl-1-(2-py- ridyl)pentyl-2-phosphonic acid, disodiumsalt

1,4-Diamino-4-methyl-1-(4-py- ridyl)pentyl-2-phosphonic acid, disodiumsalt

3-(2-Amino-4,5,7,8-tetrahydro-6H-thia- zolo[4,5-d]azepin-6-yl)propyl-phosphonic acid, disodium salt

N-Phosphonomethylglycine

N-Phosphonomethylglycine, trisodium salt

(2R,4S)-4-Phosphonomethylpipecolinic acid, trisodium salt

(2R,4S)-4-Phosphonomethyl- pipecolinamide, disodium salt

N-Phosphonomethylglycine (Aldrich, see NC-1769)

N-Phosphonomethylglycine, trisodium salt (see NC1770, prepared fromNC1781)

3-[6-Methoxy-2-(1,2,3,4-tetrahydro- isoquinolinyl)]propylphosphonicacid, disodium salt

3-[8-Methoxy-2-(1,2,3,4-tetrahydro- isoquinolinyl)]propylphosphonicacid, disodium salt

3-[2-(3-Methoxycarbonyl-1,2,3,4-tetra- hydroisoquinolinyl)]-propyl-phosphonic acid disodium salt

2-(3-Phosphonopropyl)-1,2,3,4-tetra- hydro-9H-pyrido[3,4-b]indole,disodium salt

Bisphosphonates Name Structure Pamidronic acid(3-Aminopropyl-1-hydroxy-propane-1,1-bisphosphonic acid)

3-Amino-1-hydroxypropane-1,1-bis- phosphonic acid, tetrasodium salt

1-Amino-3-sulfopropane-1,1-bis- phosphonic acid

1-Amino-3-sulfopropane-1,1-bis- phosphonic acid, pentasodium salt

1,3-Diaminopropane-1,1-bisphosphonic acid, tetrasodium salt

1-Amino-3-dimethylaminopropane-1,1-bis- phosphonic acid, tetrasodiumsalt

3-Dimethylamino-1-hydroxypropane-1,1-bis- phosphonic acid, tetrasodiumsalt

1-Hydroxy-3-(methylphenylamino)-pro- pane-1,1-bisphosphonic acid,tetrasodium salt

1-Amino-3-(methylphenylamino)propane-1,1-bis- phosphonic acid,tetrasodium salt

Ibandronic acid, tetrasodium salt (1-Hydroxy-3-(methylpentylamino)-pro-pane-1,1-bisphosphonic acid, tetrasodium salt)

1-Amino-3-(methylpentylamino)propane-1,1-bis- phosphonic acid,tetrasodium salt

1-Amino-3-(1-benzimidazolyl)propane-1,1-bis- phosphonic acid

1-Amino-3-(1-benzimidazolyl)propane-1,1-bis- phosphonic acid,tetrasodium salt

3-Aminopropane-1,1-bisphosphonic acid, tetrasodium salt

(dl)-3-Aminobutane-1,1-bisphosphonic acid, tetrasodium salt

(dl)-3-Aminopentane-1,1-bisphosphonic acid, tetrasodium salt

(dl)-3-Aminohexane-1,1-bisphosphonic acid, tetrasodium salt

(dl)-3-Aminoheptane-1,1-bisphosphonic acid, tetrasodium salt

(dl)-3-Aminooctane-1,1-bisphosphonic acid, tetrasodium salt

(dl)-3-Amino-4-methylpentane-1,1-bis- phosphonic acid, tetrasodium salt

(dl)-3-Amino-3-methylbutane-1,1-bis- phosphonic acid, tetrasodium salt

(dl)-3-Amino-3-phenylpropane-1,1-bis- phosphonic acid, tetrasodium salt

(dl)-3-Amino-4-phenylbutane-1,1-bis- phosphonic acid, tetrasodium salt

(dl)-3-Amino-4-phenylpentane-1,1-bis- phosphonic acid, tetrasodium salt

(dl)-3-Amino-3-phenylbutane-1,1-bis- phosphonic acid, tetrasodium salt

(dl)-2-(2-Amino-1,2,3,4-tetra- hydronaphthalenyl)ethane-1,1-bis-phosphonic acid, tetrasodium salt

2-(1-Aminocyclohexyl)ethane-1,1-bis- phosphonic acid, tetrasodium salt

2-(2-Amino-4,4-bisphosphonobutoxy)-tetra- hydropyran, tetrasodium salt

(dl)-3-Amino-4-hydroxybutane-1,1-bis- phosphonic acid, tetrasodium salt

(S)-Hydroxy(2-pyrrolidinyl)methane- bisphosphonic acid tetrasodium salt

Hydroxy[(2S,4R)-4-hydroxy-2-pyr- rolidinyl]methanebisphosphonic acidtetrasodium salt

2-Amino-1-hydroxyethane-1,1-bis- phosphonic acid, tetrasodium salt

1,2-Diaminoethane-1,1-bisphonponic acid, tetrasodium salt

4-Amino-1-hydroxybutane-1,1-bis- phosphonic acid, tetrasodium salt

1,4-Diaminobutane-1,1-bisphosphonic acid, tetrasodium salt

5-Amino-1-hydroxypentane-1,1-bis- phosphonic acid, tetrasodium salt

1,5-Diaminopentane-1,1-bisphosphonic acid, tetrasodium salt

(S)-2-Amino-1-hydroxypropane-1,1-bis- phosphonic acid, tetrasodium salt

(S)-2-Amino-1-hydroxybutane-1,1-bis- phosphonic acid, tetrasodium salt

(S)-2-Amino-1-hydroxy-3-methylbutane-1,1-bis- phosphonic acid,tetrasodium salt

(S)-2-Amino-1-hydroxy-3-phenylpropane-1,1-bis- phosphonic acid,tetrasodium salt

(S)-2-Amino-1,3-dihydroxypropane-1,1-bis- phosphonic acid, tetrasodiumsalt

(S)-2,3-Diamino-1-hydroxypropane-1,1-bis- phosphonic acid, tetrasodiumsalt

(dl)-3-Amino-1-hydroxy-3-phenyl- propane-1,1-bisphosphonic acid,tetrasodium salt

(S)-3-Amino-2-(4-chlorophenyl)-1-hy- droxypropane-1,1-bisphosphonicacid, tetrasodium salt

(S)-2-Amino-3-(4-aminophenyl)-1-hy- droxypropane-1,1-bisphosphonic acid,tetrasodium salt

Phosphonocarboxylate Derivatives Name Structure Phosphonoaceticacid(fosfonet)

Phosphonoformic acid, trisodium salt

Diethylphosphonoacetic acid

2-Carboxyethylphosphonic acid HO₂CCH₂CH₂PO₃H₂(dl)-2-Amino-3-phosphonopropanoic acid

(dl)-2-Amino-5-phosphonopentanoic acid

Phosphonoacetic acid(See NC-769) HO₂CCH₂PO₃H₂(S)-2-Amino-2-methyl-4-phos- phonobutanoic acid

D-(−)-2-Amino-4-phosphonobutanoic acid

L-(+)-2-Amino-4-phosphonobutanoic acid

D-(−)-2-Amino-7-phosphonoheptanoic acid

L-(+)-2-Amino-7-phosphonoheptanoic acid

D-(−)-2-Amino-6-phosphonohexanoic acid

L-(+)-2-Amino-6-phosphonohexanoic acid

D-(−)-2-Amino-4-phosphonopentanoic acid

L-(+)-2-Amino-4-phosphonopentanoic acid

D-(−)-2-Amino-3-phosphonopropanoic acid

L-(+)-2-Amino-3-phosphonopropanoic acid

(R)-(−)-3-(2-Carboxypiperazin-4-yl)-pro- pyl-1-phosphonic acid(D-CPP)

L-4-[Difluoro(phosphono)methyl)]-phenyl- alanine

(R,E)-4-(3-Phosphonoprop-2-en- yl)piperazine-2-carboxylic acid

trans-L-4-Phosphonomethylproline, trisodium salt

cis-L-4-Phosphonomethylproline, trisodium salt

N,N-Diethylphosphonoacetamide, disodium salt

N-Cyclohexylphosphonoacetamide, disodium salt

Phosphonoacetic hydrazide, disodium salt

N-Hydroxyphosphonoacetamide, disodium salt

N-Phosphonoacetyl-L-alanine, trisodium salt

N-Phosphonoacetyl-L-glycine, trisodium salt

N-(Phosphonoactyl)-L-asparagine-L-glycine, tetrasodium salt

N-Phosphonomethylglycine

N-Phosphonomethylglycine, trisodium salt

2-Phosphonomethylglutaric acid, tetrasodium salt

2-Phosphonomethylsuccinic acid, tetrasodium salt

(2R,4S)-4-Phosphonomethylpipecolinic acid, trisodium salt

(2R,4S)-4-Phosphonomethyl- pipecolinamide, disodium salt

N-Phosphonomethylglycine (Aldrich, see NC-1769)

N-Phosphonomethylglycine, trisodium salt (see NC1770, prepared fromNC1781)

3-[2-(3-Methoxycarbonyl-1,2,3,4-tetra- hydroisoquinolinyl)]-pro-pylphosphonic acid disodium salt

Phosphonate derivative Name Structure3-[2-(1,2,3,4-Tetrahydroisoquinolinyl)]-1-pro- panephosphonic acid,disodium salt

Propylphosphonic acid CH₃CH₂CH₂PO₃H₂ Ethylphosphonic acid CH₃CH₂PO₃H₂Methylphosphonic acid CH₃PO₃H₂ tert-Butylphosphonic acid (CH₃)₃CPO₃H₂Phenylphosphonic acid

3-Aminopropylphosphonic acid NH₂CH₂CH₂CH₂PO₃H₂ (1-Aminopropyl)phosphonicacid

Diethyl phosphoramidate

3-Aminopropyl(methyl)phosphinic acid, hydrochloride

4-Amino-1-butylphosphonic acid, disodium salt

1-(3-Phosphonopropyl)-benzimidazole, disodium salt

3-Dimethylamino-1-propylphosphonic acid, disodium salt

Diphenylamine-4-phosphonic acid, disodium salt

3-Amino-butylphosphonic acid, disodium salt

3-Amino-pentylphosphonic acid, disodium salt

3-Amino-hexylphosphonic acid, disodium salt

3-Amino-heptylphosphonic acid, disodium salt

3-Amino-octylphophonic acid, disodium salt

3-Amino-4-methyl-pentylphosphonic acid, disodium salt

3-Amino-3-methyl-butylphosphonic acid, disodium salt

3-Amino-3-phenyl-propylphosphonic acid, disodium salt

3-Amino-4-phenyl-butylphosphonic acid, disodium salt

3-Amino-4-phenyl-pentylphosphonic acid, disodium salt

3-Amino-3-phenyl-butylphosphonic acid, disodium salt

2-Amino-2-(2-phosphonoethyl)-1,3,4-tri- hydronaphthalene, disodium salt

1-Amino-1-(2-phosphonoethyl)-cyclo- hexane, disodium salt

2-(2-Amino-4-phos- phonobutoxy)tetrahydropyran

3-Amino-4-hydroxy-butylphosphonic acid, disodium salt

3-Phosphonopropanesulfonic acid, trisodium salt

Diethyl 2-pyrrolidinylphosphonate

2-Pyrrolidinylphosphonic acid, disodium salt

1,1-Dioxo-2-(3-phosphonopropyl)-iso- thiazoline, disodium salt

2-Deoxy-2-phosphonoacetylamino-D-glucose

3-Hydroxy-3-(2-pyridyl)propenyl-2-phos- phonic acid, disodium salt

3-Hydroxy-3-(3-pyridyl)propenyl-2-phos- phonic acid, disodium salt

3-Hydroxy-3-(4-pyridyl)propenyl-2-phos- phonic acid, disodium salt

3-Amino-3-(2-pyridyl)propenyl-2-phos- phonic acid, disodium salt

3-Amino-3-(3-pyridyl)propenyl-2-phos- phonic acid, disodium salt

3-Amino-3-(4-pyridyl)propenyl-2-phos- phonic acid, disodium salt

1,4-Diamino-1-(3-pyridyl)butyl-2-phos- phonic acid, disodium salt

1,4-Diamino-4-methyl-1-(3-py- ridyl)pentyl-2-phosphonic acid, disodiumsalt

1,4-Diamino-4-methyl-1-(2-py- ridyl)pentyl-2-phosphonic acid, disodiumsalt

1,4-Diamino-4-methyl-1-(4-py- ridyl)pentyl-2-phosphonic acid, disodiumsalt

3-(2-Amino-4,5,7,8-tetrahydro-6H-thia- zolo[4,5-d]azepin-6-yl)propyl-phosphonic acid, disodium salt

3-[6-Methoxy-2-(1,2,3,4-tetrahydro- isoquinolinyl)]propylphosphonicacid, disodium salt

3-[8-Methoxy-2-(1,2,3,4-tetrahydro- isoquinolinyl)]propylphosphonicacid, disodium salt

2-(3-Phosphonopropyl)-1,2,3,4-tetra- hydro-9H-pyrido[3,4-b]indole,disodium salt

Phosphono Carbohydrates Name Structure 2-Deoxy-2-phosphonoacetyl-amino-D-glucose

2-Deoxy-2-thiophosphono- acetylamino-D-glucose

β-D-Glucopyranosylmethyl- phosphonic acid, disodium salt

α-D-Glucopyranosylmethyl- phosphonic acid, disodium salt

6-Deoxy-6-C-phosphonomethyl- D-glucono-δ-lactone, disodium salt

6-Deoxy-6-C-phosphonomethyl- D-glucose, disodium salt

4-Deoxy-4-C-phosphonomethyl- D-glucose, disodium salt

3-Deoxy-3-C-phosphonomethyl- D-glucose, disodium salt

1-Deoxy-N-phosphonoacetyl- nojirimycin, disodium salt

(1,5-Dideoxy-1,5-imino- α-D-glucopyranosyl)methyl phosphonic acid,disodium salt

1,6-Dideoxy-6-C-phosphono- methyl-nojirimycin, disodium salt

Thiophosphonate Derivatives Name Structure Thiophosphonoformic acid,trisodium salt

Thiophosphonoacetic acid

Thiophosphonoacetic acid, trisodium salt

Thiophosphonoacetic acid, triethyl ester

Chloro(thiophosphono)acetic acid, trisodium salt

Dichloro(thiophosphono)acetic acid, trisodium salt

Thiophosphonomethylthiophosphonic acid, tetrasodium salt

Phenylthiophosphinomethylthiophosphonic acid, trisodium salt

3-[2-(1,2,3,4-Tetrahydroisoquinolinyl)]-1-propanethiophosphonic acid,disodium salt

Propylthiophosphonic acid

Ethylthiophosphonic acid

Methylthiophosphonic acid

tert-Butylthiophosphonic acid

2-Carboxyethylthiophosphonic acid

Phenylthiophosphonic acid

3-Aminopropylthiophosphonic acid

(dl)-2-Amino-3-thiophosphonopropionic acid

(1-Aminopropyl)thiophosphonic acid

(dl)-2-Amino-5-thiophosphonopentanoic acid

(S)-2-Amino-2-methyl-4-thiophosphonobutanoic acid

D-2-Amino-4-thiophosphonobutanoic acid

L-2-Amino-4-thiophosphonobutanoic acid

D-2-Amino-7-thiophosphonoheptanoic acid

L-2-Amino-7-thiophosphonoheptanoic acid

D-2-Amino-6-thiophosphonohexanoic acid

L-2-Amino-6-thiophosphonohexanoic acid

D-2-Amino-4-thiophosphonopentanoic acid

L-2-Amino-4-thiophosphonopentanoic acid

D-2-Amino-3-thiophosphonopropionic acid

L-2-Amino-3-thiophosphonopropionic acid

3-Aminopropyl(methyl)thiophosphinic acid, hydrochloride

(R)-3-(2-Carboxypiperazin-4-yl)-propyl-1-thiophosphonic acid

L-4-[Difluoro(thiophosphono)methyl)]-phenylalanine

(R,E)-4-(3-Thiophosphonoprop-2-enyl)piperazine-2-carboxylic acid

trans-L-4-Thiophosphonomethylproline, trisodium salt

cis-L-4-Thiophosphonomethylproline, trisodium salt

4-Amino-1-butylthiophosphonic acid, disodium salt

1-(3-Thiophosphonopropyl)-benzimidazole, disodium salt

3-Dimethylamino-1-propylthiophosphonic acid, disodium salt

N,N-Diethylthiophosphonoacetamide, disodium salt

Diphenylamine-4-thiophosphonic acid, disodium salt

Selenophosphonoformic acid, trisodium salt

Selenophosphonoacetic acid, trisodium salt

D-2-Amino-3-selenophosphonopropanoic acid

L-2-Amino-3-selenophosphonopropanoic acid

D-2-Amino-4-selenophosphonobutanoic acid

L-2-Amino-4-selenophosphonobutanoic acid

N-Cyclohexylthiophosphonoacetamide, disodium salt

N-Cyclohexylselenophosphonoacetamide, disodium salt

N-Hydroxythiophosphonoacetamide, disodium salt

Thiophosphonoacetic hydrazide, disodium salt

N-Thiophosphonoacetyl-L-alanine, trisodium salt

N-Thiophosphonoacetyl-L-glycine, trisodium salt

N-(Thiophosphonoactyl)-L-asparagine-L-glycine, tetrasodium salt

(s)-2-Pyrrolidinemethylthiophosphonic acid, disodium salt

1,1-Dioxo-2-(3-thiophosphonopropyl)-isothiazolidine, disodium salt

2-Deoxy-2-thiophosphonoacetylamino-D-glucose

3-(2-Amino-4,5,7,8-tetrahydro-6H-thiazolo[4,5-d]azepin-6-yl)propyl-thiophosphonic acid, disodium salt

1,1-Dioxo-2-(3-thiophosphonopropyl)-isothiazolidine, disodium salt

2-Deoxy-2-thiophosphonoacetylamino-D-glucose

3-(2-Amino-4,5,7,8-tetrahydro-6H-thiazolo[4,5-d]azepin-6-yl)propyl-thiophosphonic acid, disodium salt

1. A method of treating a condition associated with aglycosaminoglycan-associated molecular interaction in a subject,comprising administering to a subject a therapeutically effective amountof a therapeutic compound for modulating saidglycosaminoglycan-associated molecular interaction, said therapeuticcompound having the formula:Q

Y⁻X⁺]_(n) wherein Y⁻ is an anionic group at physiological pH; Q is acarrier molecule; X⁺ is a cationic group; and n is an integer selectedsuch that the biodistribution of said therapeutic compound for anintended target site is not prevented while maintaining activity of thetherapeutic compound, or a pharmaceutically acceptable salt or esterthereof, such that said glycosaminoglycan-associated molecularinteraction is modulated and said condition is treated.